High frequency plant regeneration from rice protoplasts by novel nurse culture methods

Junko Kyozuka, Yasuyuki Hayashi, Ko Shimamoto

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196 Citations (Scopus)


Novel nurse culture methods have been developed for plant regeneration from protoplasts of rice (Oryza sativa). The nurse culture methods use the agarose-bead type culture in combination with actively growing nurse cells that are either in the liquid part of the culture or inside a culture plate insert placed in the centre of the dish. Protoplasts isolated from either primary seed calluses or suspension cultures of various callus origins, divided and formed colonies with a frequency of up to 10% depending on the protoplast source and the genotype. The presence of nurse cells was absolutely required for the induction of protoplast division. Plants were regenerated from protoplast-derived calluses of five tested cultivars with a frequency of 17%-50%. Close examination of the plant regeneration process suggested that plants are regenerated through somatic embryogenesis from protoplast-derived calluses. Over 300 protoplast-derived plants were transferred to either pots or the field and are being examined for karyotypic stability and various plant phenotypes.

Original languageEnglish
Pages (from-to)408-413
Number of pages6
JournalMolecular Genetics and Genomics
Issue number3
Publication statusPublished - 1987 Mar


  • Gramineae
  • Nurse culture
  • Oryza sativa
  • Protoplast
  • Somatic embryogenesis


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